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生成具有种系嵌合能力的大鼠诱导多能干细胞。

Generation of germline-competent rat induced pluripotent stem cells.

机构信息

Japan Science Technology Agency, Exploratory Research for Advanced Technology, Nakauchi Stem Cell and Organ Regeneration Project, Tokyo, Japan.

出版信息

PLoS One. 2011;6(7):e22008. doi: 10.1371/journal.pone.0022008. Epub 2011 Jul 15.

DOI:10.1371/journal.pone.0022008
PMID:21789202
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3137610/
Abstract

BACKGROUND

Recent progress in rat pluripotent stem cell technology has been remarkable. Particularly salient is the demonstration that embryonic stem cells (ESCs) in the rat (rESCs) can contribute to germline transmission, permitting generation of gene-modified rats as is now done using mouse ESCs (mESCs) or mouse induced pluripotent stem cells (iPSCs; miPSCs). However, determinations of whether rat iPSCs (riPSCs) can contribute to germ cells are not published. Here we report the germline competency of riPSCs.

METHODOLOGY/PRINCIPAL FINDINGS: We generated riPSCs by transducing three mouse reprogramming factors (Oct3/4, Klf4, and Sox2) into rat somatic cells, followed by culture in the presence of exogenous rat leukemia inhibitory factor (rLIF) and small molecules that specifically inhibit GSK3, MEK, and FGF receptor tyrosine kinases. We found that, like rESCs, our riPSCs can contribute to germline transmission. Furthermore we found, by immunostaining of testis from mouse-rat interspecific chimeras with antibody against mouse vasa homolog, that riPSCs can contribute to embryonic development with chimera formation in mice (rat-mouse interspecific chimeras) and to interspecific germlines.

CONCLUSIONS/SIGNIFICANCE: Our data clearly demonstrate that using only three reprogramming factors (Oct3/4, Klf4, and Sox2) rat somatic cells can be reprogrammed into a ground state. Our generated riPSCs exhibited germline transmission in either rat-rat intraspecific or mouse-rat interspecific chimeras.

摘要

背景

最近大鼠多能干细胞技术取得了显著进展。特别值得注意的是，已经证明大鼠胚胎干细胞（rESCs）可以参与种系传递，从而允许生成基因修饰的大鼠，就像现在使用小鼠胚胎干细胞（mESCs）或小鼠诱导多能干细胞（miPSCs；miPSCs）所做的那样。然而，关于大鼠诱导多能干细胞（riPSCs）是否能产生生殖细胞的问题尚未发表。在这里，我们报告了 riPSCs 的种系能力。

方法/主要发现：我们通过将三个小鼠重编程因子（Oct3/4、Klf4 和 Sox2）转导到大鼠体细胞中，然后在存在外源大鼠白血病抑制因子（rLIF）和专门抑制 GSK3、MEK 和 FGF 受体酪氨酸激酶的小分子的情况下培养，从而产生 riPSCs。我们发现，与 rESCs 一样，我们的 riPSCs 可以参与种系传递。此外，我们通过用针对小鼠 vasa 同源物的抗体对来自鼠-大鼠种间嵌合体的睾丸进行免疫染色发现，riPSCs 可以在小鼠中（鼠-大鼠种间嵌合体）和种间生殖系中促进胚胎发育和嵌合体形成。

结论/意义：我们的数据清楚地表明，仅使用三个重编程因子（Oct3/4、Klf4 和 Sox2），大鼠体细胞就可以被重编程为原始状态。我们生成的 riPSCs 在大鼠-大鼠种内或鼠-大鼠种间嵌合体中均表现出种系传递。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/8fffbed63ea7/pone.0022008.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/2c7144b4ed60/pone.0022008.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/857064099dbd/pone.0022008.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/7d52927ba24f/pone.0022008.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/ac4fa60bcb39/pone.0022008.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/8fffbed63ea7/pone.0022008.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/2c7144b4ed60/pone.0022008.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/857064099dbd/pone.0022008.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/7d52927ba24f/pone.0022008.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/ac4fa60bcb39/pone.0022008.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7a3/3137610/8fffbed63ea7/pone.0022008.g005.jpg

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